Functional multineuron calcium imaging(fMCI)is an optical recording technique to monitor neuron population action potentials in the spatiotemporal pattern by recording calcium signal changes in neurons.The review describes the technology of fMCI and its application prospect in neuropharmacology research.fMCI provides a kind of powerful tool to analyze various functions of brain and to research some central nervous system drug mechanism based on neural network.

Aim To investigate the neuroprotective effect of acteoside against rotenone-induced cell damage in SH-SY5Y cells and the effect of acteoside on the expression of Parkinson disease (PD)-related proteins Parkin and ?-Synuclein(?-Syn), and to discover the underlying molecular mechanism of neuroprotection by acteoside. Methods The activity of lactate dehydrogenase(LDH) was measured by spectroscopy. Expressions of Parkin and ?-Syn were studied using Western blot analysis, and the distribution of ?-Syn was analyzed using immunofluorescence technique. Results ① Acteoside (10, 20 or 40 mg?L-1) pretreatment for 6 h markedly reduced the release of LDH induced by 0.5 ?mol ?L-1 rotenone; ② Treatment with 0.5 ?mol?L-1 rotenone for 48 h led to significant Parkin cleavage and increased formation of ?-Syn protein dimer; ③ Pretreatment of SH-SY5Y cells with acteoside (10, 20 or 40 mg?L-1) for 6 h markedly reduced the cleavage of Parkin induced by 0.5 ?mol?L-1 rotenone in a concentration-dependent manner, inhibited the aggregation of ?-Syn and decreased ?-Syn-positive SH-SY5Y cells. Conclusion These findings suggest that pretreatment of acteoside has a potent neuroprotective effect against rotenone-induced SH-SY5Y cells damage and its mechanism might involve in reducing the cleavage of Parkin and inhibitng ?-Syn expression induced by rotenone.

Recently,the techniques for proteomics have made remarkable progress.They are widely used in various fields of life sciences,providing strong technical supports for relevant researches.Especially,proteomics technology has made prominent contributions to drug development and mechanism studies,magnificently improving the efficiency of discovering new drugs.This paper summarizes the classical method and new technology of proteomics.Also,its applications in drug study,including targets screening,mechanism studies,drug toxicology,researches of resistance mechanisms and clinical medical studies have been reviewed.

Objective To investigate the changes in 20S proteasome activities in the brain and spinal cord of acute and chronic morphine-dependent mice.Metbods Male ICR mice,weighing 25-30 g,were used in the study.The experiment was performed in 2 parts.In experiment Ⅰ,16 mice were randomly divided into 5 groups (n =8 each) using a random number table:control group (group C) and acute morphine dependence group (AMD group).In experiment Ⅱ,16 mice were randomly divided into 5 groups (n =8 each) using a random number table:control group (group C) and chronic morphine dependence group (CMD group).Acute morphine dependence was induced with morphine 100 mg/kg injected subcutaneously,and the mice were sacrificed 3 h later.Chronic morphine dependence was induced by increasing doses of morphine for 4 days,the initial dose of morphine was 20 mg/kg injected subcutaneously twice a day and was increased by 10 mg/kg every day,the dose of morphine was 10 mg/kg injected subcutaneously on 5th day,and then the mice were sacrificed 1 h later.In group C,the equal volume of normal saline was given instead,and the other treatments were similar to those previously described in morphine dependence groups.After the mice were sacrificed,the hippocampus,prefrontal cortex,striatum and spinalcord were isolated for determination of 20S proteasome activity,measured as chymotrypsin-like (ChT-L),trypsin-like (T-L) and peptidylglutamyl-like hydrolyzing (PGLH) activities.Results Experiment Ⅰ Compared with C group,PGLH activity in the spinal cord and T-L activity in the striatum or prefrontal cortex were significantly weakened in group AMD.There was no significant difference in 20S proteasome activity in the hippocampus between the two groups.Experiment Ⅱ Compared with C group,ChT-L and T-L activities in the spinal cord were significantly weakened,and PGLH activity in the striatum was enhanced in CMD group.There was no significant difference in 20S proteasome activity in the prefrontal cortex and hippocampus between the two groups.Conclusion 20S proteasome activity in the spinal cord and brain is weakened in acute morphine-dependent mice,20S proteasome activity in the spinal cord is weakened,20S proteasome activity in the striatum is enhanced in chronic morphine-dependent mice,these changes have specificity in terms of position and type of activity,and the changes mentioned above may be related to development of morphine dependence in mice.

Objective To explore the characteristics of brain activation when solving simple multiplication and complex multiplication tasks. Methods From June, 2010 to June, 2012, Thirteen normal subjects completed four functional magnetic resonance imaging (fMRI) ex-periments, including control tasks, visuospatial memory tasks, simple (single-digit) multiplication tasks and complex (multi-digit) multiplica-tion tasks. Results Compared with the control tasks, visuospatial memory tasks activated the bilateral occipital lobe, the right precuneus and superior parietal lobe;simple multiplication tasks activated the bilateral middle occipital gyri, the left superior parietal lobe, the left cingu-late gyrus, the left middle frontal gyrus and inferior frontal gyrus;complex multiplication tasks activated the right superior parietal lobe, the right inferior frontal gyrus, and the bilateral middle frontal gyri. Conclusion A right parieto-frontal network is involved in the multi-digit multiplication, which supports the containing of the spatial layout information.

Objective To investigate myostatin gene mRNA expression in the muscle tissue from patients with muscle weakness suffering from different illness.Methods The clinical data of our patients were all from the Muscular Disease Center,Department of Neurology,People' s Liberation Army General Hospital.A total of 75 patients suffered from muscular weakness were included consecutively.Skeletal muscle biopsies were obtained with informed consent from all 75 patients.The diagnosis was confirmed by two senior doctors for muscular disease according to the clinical feature,the results of electromyography,serum creatine kinase activity and histopathological examination.Among of them,21 cases were diagnosed as polymyositis,15 cases progressive muscular dystrophy,5 cases neurogenic amyotrophy,4 cases chronic muscle fiber damage,4 cases mitochondrial myopathy,4 cases lipid storage myopathy,4 cases myotonic dystrophy,3 cases muscular dystrophy in adults,2 cases dermatomyositis,and 2 cases inclusion body myositis.There were 2 cases characterized by pure high activity of creatine kinase.And the other 9 cases were diagnosed as non-neuromuscular disease.The expression of myostatin gene mRNA in muscle tissue was evaluated by reverse transcription polymerase chain reaction method,with glyceraldehyde-3-phosphate dehydrogenase as internal reference.Results The expression of myostatin gene mRNA was detected in 63 patients,but not in other 12 cases,and the percentage of positive expression was 84％.The expression index was with great variation,from 0 to 3.52.In positive cohort,the index was correlated positively with the duration of disease (r =0.236,P =0.041).The activities of creatine kinase in positive expression cohort were higher than that of negative one,but nonsignificantly.Conclusion The expression of myostatin gene mRNA in muscle tissue may not correlate to the entity of atrophic muscular disease because of its great variation.

Proteomics has been applied in a wide range of biomedical research.However,the application of proteomics in studying the molecular mechanism of morphine dependence is only at a preliminary stage.This article introduced the application of proteomics techniques in the study of the molecular mechanism of morphine dependence and the discovery of several potential molecular markers of morphine dependence,which affirmed the importance and potential of proteomics in this research area.Also,it was pointed out that the major tasks of current proteomic study of morphine dependence should include establishing animal and cell models of morphine dependence,selecting appropriate sample source and improving proteomics techniques,so that proteomics can serve as a new approach in the study of morphine dependence to discover new therapeutic targets.

Aim To investigate whether acteoside can protect the dopaminergic(neurons,)(SH-SY5Y)(cells,) from rotenone-induced apoptosis.Methods Cell viability was analyzed by MTT assay.The neurons were stained with the fluorescent dye,Hoechst 33342,to analyze the nuclear change.Flow cytometry was used to determine neuronal apoptotic peak quantitatively.The level of intracellular reactive oxygen species(ROS) was monitored using the fluorescent probe 2′,7′-dichlorofluorescin-diacetate(DCFH-DA).Results ①SH-SY5Y cells treated with 0.5 ?mol ?L~(-1) rotenone for 48 h had a significantly decrease on cell viability compared with control,and the percentage of apoptosis was increased to 47.39%.The cell bodies of most cells shrinked and the dendrites of most cells became shorten or broken.Meanwhile,the shrinkage,condensation and cleavage of nuclei in most cells could be observed.The level of intracellular oxygen species was enhanced.② The preincubation with acteoside(10,20 or 40 mg?L~(-1)),one of the phenylethanoids isolated from Cistanche salsa,for 6 h enhanced the cell viability,and improved the rotenone-induced cell morphological change.Moreover,the percentage of apoptosis was significantly decreased to 25.87%,23.97% and 10.45% in a dose-dependent manner.The enhancement of ROS induced by rotenone was inhibited by 20 mg?L~(-1) acteoside.Conclusion These results demonstrate that acteoside can protect SH-SYSY cells against rotenone-induced apoptosis.The neuroprotective effect might be related with the function that acteoside can reduce the level of ROS.

Objective:To evaluate the value of color Doppler ultrasound combined with ultrasonic elastography for curative effect of neoadjuvant chemotherapy (NAC) for breast cancer.Methods: 100 patients with breast cancer who has received NAC were divided intoobservation group (50 cases, underwent examination of color Doppler ultrasound scanner combined with ultrasonic elastography) and control group (50 cases, only underwent examination of color Doppler ultrasound scanner) according to the different examination methods. The accuracy, sensitivity and specificity of differential diagnosis methods for curative effect of NAC between the two groups were compared as above data.Results: The accuracy, sensitivity and specificity of differential diagnosis for curative effect of NAC in observation group were 88.00％, 90.00％ and 86.00％, respectively. While them of control group were 78.00％, 80.00％ and 76.00％, respectively. And the differences of these indicators between the two groups were statistically significant (x2=2.01,x2=3.24,x2=3.45, P<0.05).Conclusions: Color Doppler ultrasound combined with ultrasound elastography can diagnose the curative effect of NAC for patients with breast cancer from two aspects which includes tissue hardness and blood supply, and it can achieve more diagnostic accuracy compared with only using simple color Doppler ultrasound.

Objective: To assess the effects of caffeic acid (CA) on MPP + induced cerebellar granule neurons (CGNs) apoptosis. Methods: CGNs were pretreated with caffeic acid at 55, 110 and 220 ?mol/L for 6 h, then treated with 100 ?mol/L MPP + for 24 h (concentration effect relationship). In addition CGNs were pretreated with caffeic acid at 110 ?mol/L for 0 h, 6 h, 12 h, and 24 h, respectively, then treated with 100 ?mol/L MPP + for 24 h (time response relationship). Besides, after treatment with MPP + for 24 h, CGNs were incubated with caffeic acid at 55, 110 and 220 ?mol/L,respectively. Cell viability was determined by 3 (4,5 dimethylthiazol 2 yl) 2,5 diphenyltetrazolium bromide (MTT) assay and caspase 3 activity was assayed by caspase 3 fluorometric assay kit. Results: MTT assay revealed that caffeic acid significantly inhibited cell viability decrease induced by MPP +, and caspase 3 fluorometric assay showed that caffeic acid efficiently suppressed caspase 3 activation in CGNs induced by MPP +. Conclusion: Caffeic acid (CA) can significantly protect CGNs from apoptosis induced by MPP + and may provide a useful therapeutic strategy for the treatment of Parkinson's disease.